RT Journal Article SR Electronic T1 SIRVs: Spike-In RNA Variants as External Isoform Controls in RNA-Sequencing JF bioRxiv FD Cold Spring Harbor Laboratory SP 080747 DO 10.1101/080747 A1 Lukas Paul A1 Petra Kubala A1 Gudrun Horner A1 Michael Ante A1 Igor Holländer A1 Seitz Alexander A1 Torsten Reda YR 2016 UL http://biorxiv.org/content/early/2016/10/13/080747.abstract AB Spike-In RNA variants (SIRVs) enable for the first time the validation of RNA sequencing workflows using external isoform transcript controls. 69 transcripts, derived from seven human model genes, cover the eukaryotic transcriptome complexity of start- and end-site variations, alternative splicing, overlapping genes, and antisense transcription in a condensed format. Reference RNA samples were spiked with SIRV mixes, sequenced, and exemplarily four data evaluation pipelines were challenged to account for biases introduced by the RNA-Seq workflow. The deviations of the respective isoform quantifications from the known inputs allow to determine the comparability of sequencing experiments and to extrapolate to which degree alterations in an RNA-Seq workflow affect gene expression measurements. The SIRVs as external isoform controls are an important gauge for inter-experimental comparability and a modular spike-in contribution to clear the way for diagnostic RNA-Seq applications.